BUGSS:Build-a-BUG:2-3

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=== Polymerase chain reaction (PCR) protocol ===
=== Polymerase chain reaction (PCR) protocol ===
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* Mixture
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* Assemble PCR mixture
** PCR master mix - 45.0 μL
** PCR master mix - 45.0 μL
*** Nuclease-free water -35.6 μL
*** Nuclease-free water -35.6 μL
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*** ''Taq'' DNA polymerase - 0.5 μL
*** ''Taq'' DNA polymerase - 0.5 μL
** Template DNA (3002) - 5.0 μL
** Template DNA (3002) - 5.0 μL
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* Thermal cycler program - 30 cycles
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* Amplify in thermal cycler (30 cycles)
** Melt at 95°C for ? sec
** Melt at 95°C for ? sec
** Anneal at ?°C for ? sec
** Anneal at ?°C for ? sec
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* [https://www.neb.com/products/m0273-taq-dna-polymerase-with-standard-taq-buffer ''Taq'' DNA polymerase]
* [https://www.neb.com/products/m0273-taq-dna-polymerase-with-standard-taq-buffer ''Taq'' DNA polymerase]
* [http://www.promega.com/products/pcr/routine-pcr/dntp-mix/ Deoxynucleotide triphosphate (dNTP) mix]
* [http://www.promega.com/products/pcr/routine-pcr/dntp-mix/ Deoxynucleotide triphosphate (dNTP) mix]
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== Notes ==
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*
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=== Whiteboard ===
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[[Image:BAB2-2_Agenda.jpg|thumb|left|760px|alt=Build-a-BUG:2-2 agenda|Build-a-BUG:2-2 agenda]]
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[[Image:BAB2-2_BioBricks.jpg|thumb|left|760px|alt=BioBrick parts with sizes and restriction enzymes|BioBrick parts with sizes and restriction enzymes]]
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Revision as of 15:28, 18 March 2013

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Contents

Build-a-BUG: Series 2, Session 3

Saturday, March 9, 2013
Noon to 4:00 PM

This is the third Build-A-BUG workshop in a series of five on yeast. You will learn more about molecular cloning and continue to hone your basic lab techniques, while continuing either your own or somebody else's mating type detector project.

Background reading

Polymerase chain reaction (PCR) protocol

  • Assemble PCR mixture
    • PCR master mix - 45.0 μL
      • Nuclease-free water -35.6 μL
      • 10X PCR buffer - 5.0 μL
      • Forward primer (VF2) - 1.6 μL
      • Reverse primer (VR) - 1.3 μL
      • Deoxynucleotide Triphosphate (dNTP) Mix - 1.0 μL
      • Taq DNA polymerase - 0.5 μL
    • Template DNA (3002) - 5.0 μL
  • Amplify in thermal cycler (30 cycles)
    • Melt at 95°C for ? sec
    • Anneal at ?°C for ? sec
    • Extend at 72°C for ? sec

Materials

Notes

Whiteboard

Build-a-BUG:2-2 agenda
Build-a-BUG:2-2 agenda
BioBrick parts with sizes and restriction enzymes
BioBrick parts with sizes and restriction enzymes
Personal tools